Identifying novel anti-infectives by high through-put screening in whole animals

通过对整体动物进行高通量筛选来鉴定新型抗感染药物

• 批准号: 7939581
• 负责人: Frederick M Ausubel
• 金额: $ 92.68万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-28 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939581
• 关键词: Acinetobacter; Adhesions; Animals; Anti-Infective Agents; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacteria; Bypass; Caenorhabditis elegans; Chemicals; Disease; Drosophila genus; Drosophila melanogaster; Drug Delivery Systems; Enterobacter; Epithelial; Exhibits; Funding; Generations; Goals; Gram-Negative Bacteria; Health; Human; Human body; Immune; Incidence; Infection; Klebsiella; Knowledge; Life; Microbe; Microbial Biofilms; Modeling; Molecular; Molecular Target; Multi-Drug Resistance; Nematoda; Parasites; Pathway interactions; Pharmaceutical Preparations; Physiology; Poison; Process; Pseudomonas aeruginosa; Resistance development; Screening procedure; Signal Transduction; Speed; Testing; Tissues; Toxic effect; Virulence; Virus; Work; antimicrobial drug; drug discovery; efficacy testing; fungus; high throughput screening; immunoregulation; in vivo; innovation; killings; microbial; microorganism; mouse model; next generation; novel; pathogen; prevent; public health relevance; small molecule

DESCRIPTION (provided by applicant): Continuously emerging new and hard-to-treat microbes, and the growing incidence of multi-drug resistant infections pose formidable challenges to human health. Innovative approaches are urgently needed to speed up the discovery of new anti-infectives. Our aim is to achieve a paradigm shift in antimicrobial drug discovery by finding next generation anti-infectives that prevent disease by blocking pathogen adaptation to host physiology. To this end we propose using whole live animals for high throughput screening of small molecules. We have developed infection models in the nematode Caenorhabditis elegans that can be used to identify drugs that cure otherwise lethal infections. High throughput screening of nematodes in 384-well plates is followed by secondary screening in a more highly evolved model host, the fruit fly Drosophila melanogaster, increasing the likelihood of isolating drugs that will work in humans. Our approach is applicable to many different classes of microorganisms, including bacteria, viruses, fungi and parasites. It has several advantages over traditional drug discovery: (i) In addition to identifying conventional antibiotics, it will uncover entirely new classes of anti-infectives that only exhibit in vivo activity. Examples are "virulence blockers" and "immune escape blockers". (ii) Our approach is unbiased and requires no prior knowledge of potential drug targets or pathways. (iii) It bypasses the current bottleneck of toxicity/efficacy testing by automatically eliminating toxic compounds (because they would kill the nematodes), yielding quality hits with in vivo activity. (iv) It will identify compounds that prevent or mitigate microbial resistance development, or can be combined with antibiotic therapy, thereby increasing antibiotic efficacy. We predict that our approach can identify compounds that inhibit diverse aspects of virulence: (i) adhesion and colonization, (ii) epithelial barrier disruption, (iii) deep tissue invasion, (iv) biofilm formation, (v) avoidance of immune recognition, and (vi) modulation of immune signaling. Some of the molecular mechanisms underlying these processes are conserved across bacterial species. To establish proof-of-principle, we seek funding for discovering new anti-infectives against Pseudomonas aeruginosa, one of several gram-negative bacteria that have recently emerged in a multi-drug resistant form for which efficient antibiotics are either limited or not available. We plan to screen a large number of chemical compounds (250,000) to maximize the discovery of new classes of anti-infectives. Promising compounds will undergo characterization, efficacy testing in other gram-negative bacteria (Klebsiella, Acinetobacter, Enterobacter) and testing in mouse models of infection. For highly promising candidates we will attempt molecular target identification. PUBLIC HEALTH RELEVANCE: Microbes that cause disease are becoming resistant to antibiotics faster than we can find new ones, making many common infections untreatable and life threatening. The goal of our project is to find a way to identify a new generation of antibiotics. Rather than simply preventing bacteria from growing, these new sophisticated drugs will prevent disease by interfering with a microbe's ability to interact with the human body.

描述（由申请人提供）：不断出现新的和难以治疗的微生物，以及抗多药感染的越来越多的发病率对人类健康构成了强大的挑战。迫切需要使用创新的方法来加快新反感染的发现。我们的目的是通过发现下一代抗感染剂通过阻止病原体适应宿主生理学来预防疾病的抗菌药物发现的范式转移。为此，我们建议使用全活物进行小分子的高吞吐量筛选。我们已经在线虫秀丽隐杆线虫中开发了感染模型，这些模型可用于鉴定治愈原本致命感染的药物。在384孔板中对线虫的高吞吐量筛查之后，在更高度进化的模型宿主，果蝇果蝇果皮果蝇中进行了次级筛查，增加了在人类中隔离药物的可能性。 我们的方法适用于许多不同类别的微生物，包括细菌，病毒，真菌和寄生虫。它比传统药物发现具有多个优势：（i）除了鉴定传统的抗生素外，它将发现仅显示体内活性的全新抗感染剂。例子是“毒力阻滞剂”和“免疫逃生阻滞剂”。 （ii）我们的方法是公正的，不需要对潜在药物靶标或途径的先验知识。 （iii）它通过自动消除有毒化合物（因为它们会杀死线虫）来绕过当前的毒性/功效测试的瓶颈，并随着体内活性而产生优质的命中。 （iv）它将确定预防或减轻微生物耐药性发展或可以与抗生素疗法结合的化合物，从而提高抗生素疗效。 我们预测我们的方法可以识别抑制毒力多种方面的化合物：（i）粘附和定殖，（ii）上皮屏障破坏，（iii）深层组织侵袭，（iv）生物膜形成，（v）避免免疫识别和（VI）免疫信号的调节。这些过程基础的某些分子机制在细菌物种之间是保守的。为了建立原理证明，我们寻求资金来发现针对铜绿假单胞菌的新抗感染剂，这是几种革兰氏阴性细菌之一，这些细菌最近以多药耐药形式出现，有效的抗生素是有限的，或者不可用的。我们计划筛选大量的化合物（250,000），以最大程度地发现新的抗感染剂。有希望的化合物将经历特征，在其他革兰氏阴性细菌（克雷伯菌，acinetobacter，entobacter）中进行功效测试以及在感染小鼠模型中的测试。对于高度有希望的候选人，我们将尝试分子靶标识别。 公共卫生相关性：引起疾病的微生物变得比我们发现的新抗生素更快，这使许多常见的感染无法治疗和威胁生命。我们项目的目标是找到一种方法来识别新一代的抗生素。这些新的复杂药物不仅可以防止细菌生长，还可以通过干扰微生物与人体相互作用的能力来预防疾病。